Phospho-ERK1/ERK2 (p44/p42 MAPK) (T202/Y204)

Image: Stimulation of ERK1/2 phosphorylation by NDMC in NG108-15 cells. (a) Immunofluorescence analysis of phospho-ERK1/2 immunoreactivity. Cells were serum-starved for 12 h and then treated with either vehicle (control), 10 M NDMC, 1 M NTI, and NTI+NDMC for 20 min, fixed and immunostained with anti-phospho-ERK1/2 antibody followed by FITC-conjugated secondary antibody. Results are representative of three similar experiments. (b) Western blot analysis of phospho-ERK1/2 immunoreactivity. Serum-starved cells were treated for 20 min with vehicle (lane 1), 10 M NDMC (lane 2), 1 M NTI (lane 3), and NTI+NDMC (lane 4). Thereafter, cell extracts were prepared and equal amounts of proteins (30 g) were loaded in each lane. Samples were subjected to immunoblotting with either anti-phospho-ERK1/2 antibody (top) or anti-ERK1 antibody (bottom). Results are representative of three similar experiments. (c) Densitometric analysis of immunoreactive phospho-ERK1/2 bands. The optical density of the phospho-ERK1/2 bands for each drug treatment was normalized to the density of the corresponding ERK1 band and is reported as percent of control. Values are the mean SEM of three experiments. *p<0.05 vs control, NS p>0.05 vs control by one-way ANOVA followed by Dunnett's test.